Remedy

ABSTRACT

The present invention relates to a therapeutic agent or prophylactic agent for a disease requiring enhancement of nerve growth factor (NGF) production upon treatment or prevention, an enhancing agent for nerve growth factor production, and a food, beverage or feed for enhancing nerve growth factor production, characterized in that each comprises as an effective ingredient a compound selected from the group consisting of soyasaponin compounds, soyasapogenin compounds, glycyrrhizin and salts thereof. In addition, the present invention relates to a novel soyasaponin compound and a novel soyasapogenin compound, having an action for enhancement of NGF production.

TECHNICAL FIELD

The present invention mainly relates to a medicament, foodstuff or thelike, utilizing the physiological action of a compound derived from anedible plant.

BACKGROUND ART

Nerve growth factor (NGF) is a protein closely related with life anddeath of nerve cells, showing actions such as an action for promotingbirth of nerve cells, an action for maintaining survival of nerve cells,an action for repairing the nerve cells upon brain damage, and an actionof recovering the functions of cranial nerves, thereby preventing agingof the brain. Therefore, there have been expected effects for preventingand treating Alzheimer's dementia, neural disorders in a complication ofdiabetes, or the like. However, since NGF has such a large molecularweight that does not pass through the blood-brain barrier, thedevelopment of an enhancing agent for NGF production that is capable ofpassing through the blood-brain barrier has been desired.

DISCLOSURE OF INVENTION

In view of the above-mentioned situation, an object of the presentinvention is to provide a medicament for a disease requiring enhancementfor NGF production upon treatment or prevention utilizing a safeenhancing agent for NGF production derived from natural product, andfoodstuff and the like containing the above compound in a high content.

Summarizing the present invention, the first invention, the secondinvention, the third invention and the fourth invention of the presentinvention relate to a novel compound represented by the followinggeneral formula (I):

wherein of the bond lines indicated by dotted lines, when A is a doublebond and B is a single bond, R₁ is —O-GlcUA-Gal-Glc, R₂ is—O-Ara-2-AcXyl, —O-Ara-3-AcXyl, —O-Ara-4-AcXyl, —O-Ara-2,3-diAcXyl,—O-Ara-2,4-diAcXyl, —O-Ara-3,4-diAcXyl or —O-Ara-3,4,6-triAcGlc, and R₃is —OH; alternatively, when A is a single bond and B is a double bond,R₁ is —OH, R₂ is —OH and R₃ is —H, wherein GlcUA is glucuronic acidresidue, Gal is galactose residue, Glc is glucose residue, Ara isarabinose residue, AcXyl is acetylated xylose residue, AcGlc isacetylated glucose residue, or a salt thereof; a therapeutic agent orprophylactic agent for a disease showing sensitivity to the abovecompound, an enhancing agent for nerve growth factor production, and afood, beverage or feed for enhancing nerve growth factor production,wherein each comprises the compound as an effective ingredient;.

In addition, the fifth invention, the sixth invention and the seventhinvention of the present invention relate to a therapeutic agent orprophylactic agent requiring enhancement for nerve growth factorproduction upon treatment or prevention, an enhancing agent for nervegrowth factor production, and a food, beverage or feed for enhancingnerve growth factor production characterized in that each comprises asan effective ingredient a compound selected from the group consisting ofsoyasaponin compounds, soyasapogenin compounds, glycyrrhizin andpharmacologically acceptable salts thereof.

In the embodiments of the fifth invention, the sixth invention and theseventh invention of the present invention, the soyasaponin compound andthe soyasapogenin compound are exemplified by the compound representedby the following formula (II):

wherein the bond lines indicated by dotted lines are a single bond or adouble bond; R₄ is —OH or —O— saccharide residue; R₅ is —OH, ═O or—O-saccharide residue; R₆ is —OH or —H.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a chart showing mass spectrum of the fraction 7 derived fromthe water extract of soybean embryo.

FIG. 2 is a chart showing ¹H-NMR spectrum of the fraction 7 derived fromthe water extract of soybean embryo.

FIG. 3 is a chart showing ¹³C-NMR spectrum of the fraction 7 derivedfrom the water extract of soybean embryo.

FIG. 4 is a chart showing mass spectrum of the fraction 9 derived fromthe water extract of soybean embryo.

FIG. 5 is a chart showing ¹H-NMR spectrum of the fraction 9 derived fromthe water extract of soybean embryo.

FIG. 6 is a chart showing ¹³C-NMR spectrum of the fraction 9 derivedfrom the water extract of soybean embryo.

FIG. 7 is a chart showing mass spectrum of the fraction 10 derived fromthe water extract of soybean embryo.

FIG. 8 is a chart showing ¹H-NMR spectrum of the fraction 10 derivedfrom the water extract of soybean embryo.

FIG. 9 is a chart showing ¹³C-NMR spectrum of the fraction 10 derivedfrom the water extract of soybean embryo.

FIG. 10 is a chart showing mass spectrum of the fraction 12 derived fromthe water extract of soybean embryo.

FIG. 11 is a chart showing ¹H-NMR spectrum of the fraction 12 derivedfrom the water extract of soybean embryo.

FIG. 12 is a chart showing ¹³C-NMR spectrum of the fraction 12 derivedfrom the water extract of soybean embryo.

FIG. 13 is a chart showing mass spectrum of the fraction 13 derived fromthe water extract of soybean embryo.

FIG. 14 is a chart showing ¹H-NMR spectrum of the fraction 13 derivedfrom the water extract of soybean embryo.

FIG. 15 is a chart showing ¹³C-NMR spectrum of the fraction 13 derivedfrom the water extract of soybean embryo.

FIG. 16 is a chart showing mass spectrum of the fraction 15 derived fromthe water extract of soybean embryo.

FIG. 17 is a chart showing ¹H-NMR spectrum of the fraction 15 derivedfrom the water extract of soybean embryo.

FIG. 18 is a chart showing ¹³C-NMR spectrum of the fraction 15 derivedfrom the water extract of soybean embryo.

FIG. 19 is a chart showing mass spectrum of the fraction 16 derived fromthe water extract of soybean embryo.

FIG. 20 is a chart showing ¹H-NMR spectrum of the fraction 16 derivedfrom the water extract of soybean embryo.

FIG. 21 is a chart showing ¹³C-NMR spectrum of the fraction 16 derivedfrom the water extract of soybean embryo.

FIG. 22 is a chart showing mass spectrum of the fraction a derived fromthe acid hydrolyzate of soybean embryo.

FIG. 23 is a chart showing ¹H-NMR spectrum of the fraction a derivedfrom the acid hydrolyzate of soybean embryo.

FIG. 24 is a chart showing ¹³C-NMR spectrum of the fraction a derivedfrom the acid hydrolyzate of soybean embryo.

BEST MODE FOR CARRYING OUT THE INVENTION

The present inventors have found that a specified saponin compound, aspecified sapogenin compound and/or a salt thereof is useful as anenhancing agent for NGF production, derived from an edible plant, andthe present invention has been accomplished thereby. According to thepresent invention, there could be provided a medicament containing as aneffective ingredient these saponin compound, sapogenin compound and/or apharmacologically acceptable salt thereof, foodstuff, a feed or thelike, each containing the saponin compound, the sapogenin compoundand/or a salt thereof in a high content.

The terms “enhancing action for NGF production” and “enhancing activityfor NGF production” as used herein each refers to enhancement of NGFproduction and a function for enhancing NGF production, and is notintended to particularly strictly distinguish in its meaning. Inaddition, the term “enhance” encompasses an embodiment in which theamount of the desired substance is increased after the action ascompared to that before the action of the effective ingredient of thepresent invention, as well as an embodiment (induction) in which thedesired substance is produced by the action of the effective ingredientof the present invention. In addition, any of the substances listed asthe effective ingredient in the present specification can be used aloneor in admixture of two or more kinds in the present invention.

The saponin compound, the sapogenin compound and/or a salt thereof(hereinafter referred to as “compound of the present invention” in somecases), usable as the effective ingredient in the present invention maybe those having an enhancing action for NGF production, which may bethose derived from natural products, or may be synthesized products orsemi-synthesized products. Those derived from natural product ispreferably those derived from an edible plant, and those derived from anedible plant are exemplified by soyasaponin compounds or soyasapogenincompounds derived from beans such as soybeans. Also, there may beexemplified glycyrrhizin which is known as a saponin derived fromglycyrrhiza.

The soybeans include four groups of saponins of A, B, E and DDMPsaponins depending upon their chemical structures. The group A saponinis a saponin in which olean-12-en-3β,21β,22β,24-tetraol (soyasapogenolA) constitutes the aglycon; the group B saponin is a saponin in whicholean-12-en-3β,22β,24triol (soyasapogenol B) constitutes the aglycon;further the group E saponin is a saponin in whicholean-12-en-3β,24-diol-22-one (soyasapogenol E) constitutes the aglycon;and the DDMP saponin is a saponin in which the group B saponinconstitutes the aglycon provided that2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one is bound at C-22position (Shokuhin to Kaihatsu (Food and Development), Vol. 34, No. 7,pages 8-11, (1999)). In other words, in the present invention, as thesoyasaponin compounds, the group A, B and E saponins and the DDMPsaponin are especially suitably used. In addition, the compound of thepresent invention is exemplified by the compound represented by theabove-mentioned general formula (II), especially preferably compounds ato n represented by the following general formula (III).

wherein the bond lines C and D indicated by dotted lines, and R₇, R₈,and R₉ are shown in Table 1. TABLE 1 C D R₇ R₈ R₉ Compound a DoubleSingle -O-GlcUA-Gal-Glc -O-Ara-2-AcXyl —OH bond bond Compound b DoubleSingle -O-GlcUA-Gal-Glc -O-Ara-3-AcXyl —OH bond bond Compound c DoubleSingle -O-GlcUA-Gal-Glc -O-Ara-4-AcXyl —OH bond bond Compound d DoubleSingle -O-GlcUA-Gal-Glc -O-Ara-2,3-diAcXyl —OH bond bond Compound eDouble Single -O-GlcUA-Gal-Glc -O-Ara-2,4-diAcXyl —OH bond bond Compoundf Double Single -O-GlcUA-Gal-Glc -O-Ara-3,4-diAcXyl —OH bond bondCompound g Double Single -O-GlcUA-Gal-Glc -O-Ara-3,4,6-triAcGlc —OH bondbond Compound h Double Single -O-GlcUA-Gal-Glc -O-Ara-2,3,4,6-tetraAcGlc—OH bond bond Compound i Double Single -O-GlcUA-Gal-Glc —OH —H bond bondCompound j Double Single -O-GlcUA-Gal-Rham —OH —H bond bond Compound kDouble Single -O-GlcUA-Gal-Glc ═O —H bond bond Compound l Double Single-O-GlcUA-Gal-Rham ═O —H bond bond Compound m Double Single-O-GlcUA-Gal-Glc -O-Ara-2,3,4-triAcXyl —H bond bond Compound n SingleDouble —OH —OH —H bond bond

Here, the above-mentioned compounds a to g and n, specifically thecompounds represented by the above-mentioned general formula (I), arecompound obtained for the first time according to the present invention,of which enhancing actions for NGF production have been elucidated. Inthe present invention, there are also provided the compound, and atherapeutic agent or prophylactic agent of a disease showing sensitivityto the compound, comprising the compound as an effective ingredient.

In addition, as the compound of the present invention, those derivedfrom soybeans and glycyrrhiza are preferable. The compound of thepresent invention is not limited in derivations thereof, as long as thecompound is a soyasaponin compound, a soyasapogenin compound orglycyrrhizin, for instance, the compound represented by theabove-mentioned general formula (II), even if the compound is notderived from soybean. In the present specification, AcXyl meansacetylated xylose residue, AcGlc means acetylated glucose residue, andRham means rhamnose residue. Also, the soyasapogenin compound meansaglycon or isomer thereof without a saccharide residue in thesoyasaponin compound.

The sugar constituting the saccharide residue may be a monosaccharide ora sugar chain, and the saccharide residue may comprise one or moresaccharides. For instance, the sugar is exemplified by, for instance,glucose, glucuronic acid, xylose, rhamnose, galactose, threose, ribose,apiose, allose, arabinose, arabinopyranose, ribulose, mannose, talose,fucose, fructose, galacturonic acid, and sugar chains constituted bythese saccharides. The number of the saccharides in the sugar chain is,but not particularly limited to, especially preferably from 2 to 5, fromthe viewpoint of exhibiting the desired effects of the presentinvention. Also, these saccharides may be subjected to a modificationsuch as acetylation or esterification.

Furthermore, the compound of the present invention can be formed into aderivative (prodrug), which can be readily hydrolyzed in the body toexhibit the desired effects, by, for instance, subjecting the compoundto esterification. The prodrug may be prepared in accordance with aknown method. Here, the derivative may be their salts.

In addition, in the compound of the present invention, as the salts,pharmacologically acceptable salts are preferable. The salts used in thepresent invention are exemplified by, for instance, alkali metal salts,alkaline earth metal salts, salts with an organic base, and the like.The pharmacologically acceptable salt used in the present invention isreferred to a salt of a compound which is substantially nontoxic againstan organism, and has an enhancing action for NGF production. The saltsof the compound as the effective ingredient of the present inventioninclude, for instance, salts with sodium, potassium, calcium, magnesium,ammonium or protonated benzathine (N,N′-di-benzylethylenediamine),choline, ethanolamine, diethanolamine, ethylenediamine, meglamine(N-methylglucamine), benethamine (N-benzylphenetylamine), piperazine ortolomethamine (2-amino-2-hydroxymethyl-1,3-propanediol).

Also, various isomers such as optical isomers, keto-enol tautomers, andgeometric isomers of the compound of the present invention can be allused in the present invention, even when each of isomers is isolated, aslong as these isomers have an enhancing activity for NGF production.Therefore, the compound of the present invention encompasses derivativesthereof, isomers thereof and salts thereof, as long as the desiredeffects of the present invention can be obtained.

If the compound of the present invention is commercially available, sucha compound can be utilized. Also, the compound of the present inventioncan be prepared. The preparation process includes a process comprisingextracting and purifying the compound from a plant to give the compoundin accordance with a conventional method.

The compound of the present invention can be prepared from a naturalproduct by combining known preparation processes. For instance, thecompound can be extracted and purified from a substance containing asoyasaponin compound, a soyasapogenin compound or a salt thereof, forinstance, a plant such as soybean. In addition, when soybean is used,soybean embryo and cotyledon are preferably used as raw materials. Asthe extraction solvent, there can be used, for instance, a hydrophilicor lipophilic solvent such as water, chloroform, an alcohol such asethanol, methanol or isopropyl alcohol, a ketone such as acetone ormethyl ethyl ketone, methyl acetate or ethyl acetate, alone or as aliquid mixture. The extraction temperature and time may be appropriatelyset as desired, and the extraction procedures may be repeated severaltimes as desired. As the purification means, a known purification meanssuch as a chemical method or physical method may be employed. Aconventionally known purification method such as gel filtration method,fractionation method with a molecular weight fractionation membrane,solvent extraction method, and various chromatographic methods using ionexchange resins and the like may be combined to concentrate, isolate orthe like the desired compound to be used as the effective ingredient.The medicament or the like of the present invention described later canbe prepared by using the concentrate of the compound, without having touse the isolated compound.

In addition, glycyrrhizin usable in the present invention is notparticularly limited. Glycyrrhizin is commercially available, or can beobtained by extracting and purifying from glycyrrhiza. The extractionand purification of glycyrrhizin can be carried out in accordance withthe method described for the above-mentioned soyasaponin compound or thelike. Also, in the present invention, the glycyrrhizin derivative can beused.

When the compound usable as the effective ingredient in the presentinvention is obtained as a synthesized product or semi-synthesizedproduct, the desired compound may be synthesized, for instance, inaccordance with a known method.

Whether or not the desired compound is obtained can be confirmed by, forinstance, determining the structure by mass spectroscopy, nuclearmagnetic resonance method or the like as described in Examples set forthbelow.

According to the present invention, there is provided a therapeuticagent or prophylactic agent for a disease requiring enhancement of NGFproduction upon treatment or prevention, comprising as the effectiveingredient the compound of the present invention as described above.

NGF is an endogenous growth factor for maintaining viability andfunctions of nerve cells, elongating nerve cells in accordance with aconcentration gradient of NGF, or the like. By enhancing the productionof NGF, the treatment or prevention of senile dementia such asAlzheimer's disease, peripheral nerve disorder, cerebrovasculardisorder, cerebral tumor, cerebral apicitis, nerve degenerative diseasecaused by head injury, diseases requiring recovery and regeneration ofnerve functions, caused by intoxication with an anesthetic or the likecan be carried out. In addition, it is useful in the treatment orprevention of amyotrophic lateral sclerosis, drug-induced peripheralnerve disorder, diabetic peripheral nerve disorder, Parkinson's disease,sensory nerve disorder, retinitis pigmentosa, macular dystrophy, and thelike. In other words, by administering or taking the medicament, or thefoodstuff or feed mentioned later of the present invention, thetreatment or prevention of the above-mentioned diseases can be carriedout.

The medicament of the present invention encompasses a therapeutic agentor prophylactic agent, characterized in that the therapeutic agent orprophylactic agent comprises as the effective ingredient the compoundrepresented by the general formula (I) and/or a pharmacologicallyacceptable salt thereof, which are obtained for the first time in thepresent invention, wherein the therapeutic agent or prophylactic agentis for a disease showing sensitivity to the above compound.

The above-mentioned therapeutic agent or prophylactic agent of thepresent invention can be prepared by combining the compound of thepresent invention, for instance, a soyasaponin compound, a soyasapogenincompound, glycyrrhizin or a pharmacologically acceptable salt thereof asan effective ingredient with a known pharmaceutical carrier to form intoa preparation.

The content of the effective ingredient in the medicament of the presentinvention cannot be absolutely determined because the content differsdepending upon the diseases to be treated or the like, use embodimentsand the like. The content is usually from 0.01 to 50% by weight or so,preferably from 0.1 to 10% by weight or so.

In general, the therapeutic agent or prophylactic agent of the presentinvention is manufactured by formulating the compound of the presentinvention with a pharmacologically acceptable liquid or solid carrier,and optionally a solvent, a dispersant, an emulsifier, a buffer, astabilizer, an excipient, a binder, a disintegrant, a lubricant, or thelike can be added thereto, so that a solid agent such as a tablet, agranule, a powder, a fine powder, or a capsule, or a liquid agent suchas a common liquid agent, a suspension agent or an emulsion agent can beformed. In addition, there can be also made into a dry product which canbe made liquid by adding an appropriate liquid vehicle before use.

The therapeutic agent or prophylactic agent of the present invention canbe administered in any of the forms of orally administered agents andnon-orally administered agents such as injections and drops.

The dose of the therapeutic agent or prophylactic agent of the presentinvention is changeable and properly set depending upon its preparationform, administration method, purpose of use, and age, body weight,symptom of a patient to which the therapeutic agent or prophylacticagent is applied. Generally, the dose of the agent as the dose for humanper day of the effective ingredient contained in the preparation isusually from 1 μg to 1 g/kg body weight, preferably from 10 μg to 200mg/kg body weight. As a matter of course, the dose varies depending uponvarious conditions, so that an amount smaller than the dose mentionedabove may be sufficient, or an amount exceeding the dose range may berequired. Also, the period of administration is not particularlylimited. The medicament of the present invention can be directly orallyadministered, or the agent can be added to any foodstuffs be taken on adaily basis.

In addition, the present invention can provide an enhancing agent forNGF production, comprising the compound of the present invention as theeffective ingredient. The enhancing agent may be the above-mentionedeffective ingredient itself, or a composition comprising theabove-mentioned effective ingredient. In the embodiment of the presentinvention, the salt as the effective ingredient is preferably apharmacologically acceptable salt. The enhancing agent for NGFproduction may be produced by, for instance, formulating theabove-mentioned effective ingredient with other ingredients which can beused for the same application as the effective ingredient, and forminginto a form of reagent usually used according to the above-mentionedprocess for producing the therapeutic agent or prophylactic agent. Thecontent of the above-mentioned effective ingredient in the enhancingagent is not particularly limited, as long as the content is in anamount so that the desired effects of the present invention can beexhibited in consideration of administration method, purpose of use, orthe like of the enhancing agent. The content of the effective ingredientof the present invention in the enhancing agent for NGF production isusually from 0.01 to 50% by weight or so, preferably from 0.1 to 10% byweight or so. Also, the amount of the enhancing agent used is notparticularly limited, as long as the desired effects of the presentinvention can be exhibited. Especially in the case where the enhancingagent is administered to a living body, the enhancing agent ispreferably used in an amount so that the effective ingredient can beadministered within the dose range of the effective ingredient for theabove-mentioned therapeutic agent or prophylactic agent. The enhancingagent for NGF production is useful for enhancement for NGF production ina disease requiring enhancement for NGF production. In addition, theenhancing agent is also useful for screening of drugs for diseasesassociated with NGF. Furthermore, the enhancing agent is useful forfunctional studies concerning NGF or physical changes in nerve cells.

Next, there is explained a food or beverage (hereinafter referred to as“foodstuff” in some cases) or a feed, each comprising theabove-mentioned effective ingredient. In the foodstuff or feed of thepresent invention, the term “comprising” encompasses the meaning ofcontaining, adding and diluting. Therefore, the foodstuff or feed of thepresent invention is a food, beverage or feed in which the compound ofthe present invention is contained, added and/or diluted. The foodstuffor the like is very useful in the amelioration of symptoms or preventionof a disease requiring enhancement for NGF production upon treatment orprevention. In addition, the foodstuff or the like is useful in themaintenance of homeostasis of a living body. The salt of the compoundusable as the effective ingredient is preferably a pharmacologicallyacceptable salt.

As used herein in the foodstuff or feed of the present invention, theterm “containing” refers to an embodiment of containing the effectiveingredient usable in the present invention in the foodstuff or the like;the term “adding” refers to an embodiment of adding the effectiveingredient usable in the present invention to a raw material for thefoodstuff or the like; and the term “diluting” refers to an embodimentof adding a raw material for the foodstuff or the like to the effectiveingredient usable in the present invention.

The process for preparing the food, beverage or feed of the presentinvention is not particularly limited. A generally used process forpreparing a food, beverage or feed can be employed, as long as theresulting food, beverage or feed may contain the compound of the presentinvention as the effective ingredient in a high content, wherein thecontent is higher than that of a usual foodstuff or feed. In otherwords, the foodstuff or feed of the present invention can be prepared byconcentrating or isolating the effective ingredient of the presentinvention from the raw materials, and formulating the effectiveingredient in a known foodstuff or feed. The concentration or isolationencompasses the synthesis of the effective ingredient. Also, when theraw materials can be treated such that the content of the effectiveingredient is increased in the raw materials, the resulting processedproduct can be directly used as the foodstuff of the present invention,as long as the raw materials may be used for edible purposes for human.For instance, the processed products include concentrated juice,vegetable juice and the like. The expression “contained . . . in a highcontent” as used herein means that the weight of the compound of thepresent invention per unit weight of the food, beverage or feed of thepresent invention is larger than the weight of the compound of thepresent invention per unit weight of the raw material, for instance,soybean, for the effective ingredient of the present invention. Also,the weight refers to the weight of the solid content.

The food or beverage of the present invention is not particularlylimited. The food or beverage includes, for instance, processedagricultural and forest products, processed stock raising products,processed marine products and the like, including processed grainproducts (for instance, processed wheat products, processed starchproducts, processed premix products, noodles, macaronis, bread, beanjam, buckwheat noodles, wheat-gluten bread, rice noodle, fen-tiao,packed rice cake and the like); processed fat and oil products (forinstance, plastic fat and oil, tempura oil, salad oil, mayonnaise,dressing and the like); processed soybean products (for instance, tofuproducts, soybean paste, fermented soybeans and the like); processedmeat products (for instance, ham, bacon, pressed ham, sausage and thelike); marine products (for instance, frozen ground fish, boiled fishpaste, tubular roll of boiled fish paste, cake of ground fish,deep-fried patty of fish paste, fish ball, sinew, fish meat ham,sausage, dried bonito, products of processed fish egg, marine cans,preserved food boiled down in soy sauce (tsukudani) and the like); milkproducts (for instance, raw material milk, cream, yogurt, butter,cheese, condensed milk, powder milk, ice cream and the like); processedvegetable and fruit products (for instance, paste, jam, pickledvegetables, fruit beverages, vegetable beverages, mixed beverages andthe like); confectionaries (for instance, chocolates, biscuits, sweetbun, cake, rice cake snacks, rice snacks and the like); alcoholbeverages (for instance, sake, Chinese liquor, wine, whiskey, Japanesedistilled liquor (shochu), vodka, brandy, gin, rum, beer, refreshingalcoholic beverages, fruit liquor, liqueur and the like); luxury drinks(for instance, green tea, tea, oolong tea, coffee, refreshing beverages,lactic acid beverages and the like); seasonings (for instance, soysauce, sauce, vinegar, sweet rice wine and the like); canned, binned orpouched foods (for instance, rice topped cooked beef and vegetable, riceboiled together with meat and vegetables in a small pot, steamed ricewith red beans, curry roux and rice, other precooked foods and thelike); semi-dry or concentrated foods (for instance, liver pastes andother spreads, soups for buckwheat noodles or wheat noodles,concentrated soups and the like); dry foods (for instance, instantnoodles, instant curry roux, instant coffee, powder juice, powder soup,instant soybean paste (miso) soup, precooked foods, precooked beverages,precooked soup and the like); frozen foods (for instance, sukiyaki,pot-steamed hotchpotch, split and grilled eel, hamburger steak,shao-mai, dumpling stuffed with minced pork, various sticks, fruitcocktails and the like); solid foods; liquid foods (for instance, soupsand the like); spices; and the like.

The content of the effective ingredient in the foodstuff of the presentinvention is not particularly limited, and the content is usually from0.01 to 50% by weight or so, preferably from 0.1 to 10% by weight or so.

The feed of the present invention is, for instance, a feed for cattle,culture fish, poultry, or pet, which means an artificial food orbeverage for an organism other than human, comprising the compound ofthe present invention. The beverage includes drinking water for cattleand the like. In addition, one embodiment of the present inventionprovides a feed additive for enhancement of NGF production, comprisingthe effective ingredient of the present invention. The additive is usedby adding to an ordinary feed or water in which an organism is immersed(water for culturing a culture fish, or the like).

The content of the effective ingredient in the feed of the presentinvention is not particularly limited, and the content is usually from0.01 to 50% by weight or so, preferably from 0.1 to 10% by weight or so.The content of the effective ingredient in the above-mentioned additiveis not particularly limited, and may be properly adjusted so as to givethe desired effects according to its purposes.

The form of the feed (including the above-mentioned additive) of thepresent invention is not particularly limited, and may be in the form ofa powder, a liquid or a solid. Here, the salt of the compound usable asthe effective ingredient is preferably a pharmacologically acceptablesalt.

In the present invention, the content of the compound of the presentinvention in the medicament, foodstuff or feed is as mentioned above.The compound is contained in a high content which is equal to or higherthan the content of the soyasaponin compound, the soyasapogenin compoundand glycyrrhizin in an ordinary foodstuff, as long as the content issuch an amount that NGF production is enhanced in a living body uponadministration, intake or the like.

No toxicity is found in the compound of the present invention, which isthe effective ingredient. For instance, no toxicity is found when amouse (male, 6-week old) is orally administered with the saponincompound, glycyrrhizin or a salt thereof usable in the present inventionat 1000 mg per kg body weight.

In addition, the present invention also provides a novel soyasaponincompound and a novel soyasapogenin compound, which are represented bythe above-mentioned general formula (I), wherein each of the compoundshas an enhancing action for NGF production. The process for thepreparation is as described above, and is described in further detail bymeans of the following Examples. The novel compounds of the presentinvention can be prepared in reference to the Examples, without beinglimited thereto.

EXAMPLES

The present invention will be more concretely described hereinbelow bymeans of the examples, without by no means limiting the scope of thepresent invention thereto.

Example 1 Fractionation of Fraction Derived from Water Extract ofSoybean Embryo

(1) Twenty-five liters of distilled water was added to 9.6 kg of drysoybean embryo, and the soybean embryo was immersed for one hour, andthereafter disrupted with a blender. Next, thereto was added 40 litersof distilled water, and the mixture was stirred for 2 hours. Thereafter,centrifugation was carried out at 5000 g for 7 minutes, to give aprecipitate 1 and supernatant 1 (54 liters). Forty liters of distilledwater was added to the precipitate 1, and the mixture was stirred for 2hours. Thereafter, centrifugation was carried out at 5000 g for 7minutes, to give a precipitate 2 (19.7 kg) and supernatant 2 (36liters). The supernatant 1 and the supernatant 2 were mixed together, togive 90 L of a water extract of soybean embryo.

(2) Thirty-nine liters of ethanol was added to 90 liters of the waterextract of soybean embryo obtained in item (1) of Example 1, and themixture was stirred. Thereafter, centrifugation was carried out at 5000g for 7 minutes, to give a precipitate 3 and supernatant 3 (90 liters).The supernatant 3 was concentrated to a volume of 18 liters with arotary evaporator, an equal volume of ethanol was added thereto, and themixture was stirred. Thereafter, centrifugation was carried out at 5000g for 7 minutes, to give a precipitate 4 and supernatant 4 (15 liters).The supernatant 4 was concentrated to give 1.7 liters of a concentrateof water extract of soybean embryo.

(3) A mixture prepared by adding 500 mL of distilled water to 110 mL ofthe concentrate of water extract of soybean embryo obtained in item (2)of Example 1 was fractionated using reverse phase chromatography. Theresin used was Cosmosil 140 C18-OPN (manufactured by nakalai tesque,amount of resin: 300 mL). The elution and fractionation were carried outusing 1500 mL of distilled water, a 30% aqueous ethanol solution, a 50%aqueous ethanol solution, and 100% ethanol in that order as thedeveloping solvents, to give each of Cosmosil-eluted fractions derivedfrom the water extract of soybean embryo.

(4) The Cosmosil-30% ethanol-eluted fraction derived from the waterextract of soybean embryo obtained in item (3) of Example 1 wasconcentrated under reduced pressure. Thereafter, acetone was added tothe concentrate in an amount three times the volume of the concentrate,and the mixture was stirred, and centrifugation was carried out at 5000g for 7 minutes, to give a precipitate 5 and supernatant 5.

(5) The supernatant 5 obtained in item (4) of Example 1 was concentratedto dryness, and thereafter the residue was dissolved in ethylacetate:acetic acid:water (volume ratio, hereinafter the same)=8:3:2 (10mL). The mixture was fractionated using silica chromatography. Theconditions therefor are given below. BW-300SP (manufactured by FujiSilysia Chemical Ltd.; amount of resin: 300 mL) was used as the silicagel. The elution was carried out using ethyl acetate:aceticacid:water=8:3:2 (1000 mL) and ethanol:water=5:1 (500 mL) in that orderas the developing solvents, to give eluted fractions of a fraction 0(300 mL), a fraction 1 (200 mL), a fraction 2 (200 mL), a fraction 3(100 mL), a fraction 4 (150 mL), a fraction 5 (200 mL) and a fraction 6(120 mL) in that order. Each of the eluates was concentrated underreduced pressure, to give each of silica column fractions 0 to 6 derivedfrom the water extract of soybean embryo.

(6) The silica column fraction 5 derived from the water extract ofsoybean embryo obtained in item (5) of Example 1 was dissolved in 60 mLof distilled water. Thereafter, the solution was fractionated usingreverse phase chromatography. The conditions therefor are given below.TSK gel ODS-80Ts (21.5 mm×30 cm; manufactured by Tosoh Corporation) wasused as a column. The elution ratio of Solvent A (mixture prepared bymixing distilled water and acetonitrile in a volume ratio of 1:3) toSolvent B (mixture prepared by mixing distilled water and acetonitrilein a volume ratio of 3:1) was such that the ratio of Solvent A wasretained at 100% from 0 to 10 minutes, the ratio of Solvent B increasedlinearly from 0 to 30% from 10 to 25 minutes, the ratio of Solvent Bincreased linearly from 30 to 100% from the next 25 to 40 minutes, andthe ratio of Solvent B was retained at 100% for the subsequent 15minutes. The elution rate was 5 mL/minute, and the detection was carriedout at 215 nm. The fractions 1 to 21 derived from the water extract ofsoybean embryo were fractionated using ultraviolet absorption of aneluate as an index.

Example 2 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The mass spectrum (MS) of the fraction 7 derived from the waterextract of soybean embryo (fraction including a peak detected at aretention time of 32.4 minutes) fractionated in item (6) of Example 1was measured with a mass spectrometer (DX302, manufactured by JEOL LTD.)by FAB-MS technique. As the matrix, triethanolamine was used. As aresult, a peak of m/z 1279 (M−H)⁻ was detected. FIG. 1 shows the massspectrum of the fraction 7 derived from the water extract of soybeanembryo. In FIG. 1, the axis of abscissas is m/z value, and the axis ofordinates is relative intensity.

The structure of the fraction 7 derived from the water extract ofsoybean embryo was analyzed by measuring various NMR spectra usingnuclear magnetic resonance (NMR) spectrometer (Model AVANCE600,manufactured by Bruker Biospin K.K.). The signals of NMR are shownbelow.

¹H-NMR: sapogenol moiety; δ3.28 (1H, m, 3-H), 5.15 (1H, br-s, 12-H),3.45 (1H, m, 21-H), 3.23 (1H, br-s, 22-H), 3.14 (1H, m, 24-H), 3.85 (1H,m, 24-H) 3-O-β-D-glucronopyranosyl moiety; δ4.45 (1H, d, J=7.2 Hz,1′-H), 3.44 (1H, m, 2′-H), 3.57 (1H, m, 3′-H), 3.32 (1H, m, 4′-H), 3.65(1H, m, 5′-H) 2′-O-β-D-galactopyranosyl moiety; δ4.81 (1H, d, J=6.6 Hz,1″-H), 3.51 (1H, m, 2″-H), 3.51 (1H, m, 3″-H), 3.65 (1H, m, 4″-H), 3.38(1H, m, 5″-H), 3.50 (2H, m, 6″-H) 2″-O-β-D-glucopyranosyl moiety; δ4.38(1H, d, J=7.8 Hz, 1′″-H), 3.02 (1H, m, 2′″-H), 3.17 (1H, m, 3′″-H), 3.08(1H, m, 4′″-H), 3.14 (1H, m, 5′″-H), 3.46 (1H, m, 6′″-H), 3.73 (1H, m,6′″-H) 22-O-α-L-arabinopyranosyl moiety; δ4.13 (1H, m, 1″″-H), 3.42 (1H,m, 2″″-H), 3.42 (1H, m, 3″″-H), 3.69 (1H, m, 4″″-H), 3.35 (1H, m,5″″-H), 3.62 (1H, m, 5″″-H) 3″″-O-β-D-xylopyranosyl moiety; δ4.57 (1H,d, J=7.8 Hz, 1″″′-H), 4.52 (1H, m, 2″″′-H), 3.29 (1H, m, 3″″′-H), 3.37(1H, m, 4″″′-H), 3.09 (1H, m, 5″″′-H), 3.72 (1H, m, 5″″′-H), 2.00 (3H,s, 2″″′-CH₃)

Here, in ¹H-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of the residual proton of deutrateddimethyl sulfoxide was expressed as 2.51 ppm. FIG. 2 shows ¹H-NMRspectrum of the fraction 7 derived from the water extract of soybeanembryo. In FIG. 2, the axis of abscissas is chemical shift, and the axisof ordinates is intensity of signal.

¹³C-NMR: sapogenol moiety; δ 90.3 (3-C), 122.5 (12-C), 144.4 (13-C),74.7 (21-C), 92.0 (22-C), 63.0 (24-C) 3-O-β-D-glucronopyranosyl moiety;δ104.1 (1′-C), 79.0 (2′-C), 76.6 (3′-C), 71.9 (4′-C), 76.2 (5′-C), 171.0(6′-C) 2′-O-β-D-galactopyranosyl moiety; δ101.6 (1″-C), 82.7 (2″-C),73.4 (3″-C), 69.0 (4″-C), 75.5 (5″-C), 60.9 (6″-C)2″-O-β-D-glucopyranosyl moiety; δ105.3 (1′″-C), 75.5 (2′″-C), 76.7(3′″-C), 70.6 (4′″-C), 78.1 (5′″-C), 61.8 (6′″-C)22-O-α-L-arabinopyranosyl moiety; δ108.0 (1″″-C), 71.9 (2″″-C), 83.0(3″″-C), 68.5 (4″″-C), 67.0 (5″″-C) 3″″-O-β-D-xylopyranosyl moiety;δ103.3 (1″″′-C), 74.6 (2″″′-C), 74.7(3″″′-C), 70.2 (4″″′-C), 65.3(5″″′-C), 170.5 (2″″′-C═O), 21.8 (2″″′-CH₃)

Here, in ¹³C-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 40.2 ppm. FIG. 3 shows ¹³C-NMR spectrum of the fraction 7derived from the water extract of soybean embryo. In FIG. 3, the axis ofabscissas is chemical shift, and the axis of ordinates is intensity ofsignal.

The fraction 7 derived from the water extract of soybean embryo wasmixed with 1 N H₂SO₄ (solution having a volume ratio of dioxane andwater of 1:3), and the mixture was heated at 80° C. for 4 hours, toproduce a saccharide component and a non-saccharide component. Thenon-saccharide component was subjected to thin layer chromatography(Silica gel 60 F254 manufactured by Merck, developing solvent A:prepared by mixing chloroform and methanol in a ratio of 10:1). As aresult, a spot having the same Rf values as in Soyasapogenol A wasdetected. The saccharide component was subjected to thin layerchromatography (developing solvent B: prepared by mixing water andacetonitrile in a ratio of 3:17). As a result, arabinose, glucose,xylose and galactose were detected.

From the above analyses of the mass spectrum and NMR spectrum and theanalyses of the non-saccharide component and the saccharide componentafter acid hydrolysis performed for the fraction 7 derived from thewater extract of soybean embryo, it was identified that the fraction 7derived from the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1280). The compound is a compound a in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound a wasassayed. L-M cells (ATCC CCL-1.2) from murine fibroblasts were suspendedin an M199 medium (manufactured by Bio Whittaker) containing 0.5%bactopeptone (manufactured by Gibco) so as to have a concentration of1.5×10⁵ cells/mL. The suspension was put in a 96 well plate in an amountof 0.1 mL each well, and the cells were aseptically cultured. Afterculturing the cells for 3 days, the medium was removed therefrom, andexchanged with an M199 medium containing 0.5% bovine serum albumin(manufactured by Sigma). The compound a was added thereto, and the cellswere cultured for 20 hours. After the termination of the culture, theconcentration of NGF in the culture medium was assayed by an enzymeimmunoassay method (NGF Emax Immuno Assay System, manufactured byPromega). The added amount, as expressed in the final concentration, isas shown in Table 2. The experiment was carried out twice, and anaverage value was taken. As a result, it was clarified that the compounda has an enhancing activity for NGF production. The results are shown inTable 2. The amount of NGF production of the control was 0.739 ng/mL. Asto the control, the concentration in the culture medium was determinedin the same manner except that the compound a was not added. It is thesame as that of the following examples.

Example 3 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[3-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The mass spectrum of the fraction 9 derived from the water extractof soybean embryo (fraction including a peak detected at a retentiontime of 34.2 minutes) fractionated in item (6) of Example 1 was measuredin the same manner as in item (1) of Example 2. As the matrix,triethanolamine was used. According to mass spectrometry, a peak of m/z1279 (M−H)⁻ was detected. FIG. 4 shows the mass spectrum of the fraction9 derived from the water extract of soybean embryo. In FIG. 4, the axisof abscissas is m/z value, and the axis of ordinates is relativeintensity.

The NMR spectrum of the fraction 9 derived from the water extract ofsoybean embryo was measured in the same manner as in item (1) of Example2. The signals of NMR are shown below.

¹H-NMR: sapogenol moiety; δ3.27 (1H, m, 3-H), 5.14 (1H, br-s, 12-H),3.39 (1H, m, 21-H), 3.24 (1H, br-s, 22-H), 3.14 (1H, m, 24-H), 3.86 (1H,m, 24-H) 3-O-β-D-glucronopyranosyl moiety; δ4.45 (1H, m, 1′-H), 3.43(1H, m, 2′-H), 3.57 (1H, m, 3′-H), 3.33 (1H, t, J=9.7 Hz, 4′-H), 3.65(1H, m, 5′-H) 2′-O-β-D-galactopyranosyl moiety; δ4.80 (1H, d, J=6.6 Hz,1″-H), 3.51 (1H, m, 2″-H), 3.51 (1H, m, 3″-H), 3.65 (1H, m, 4″-H), 3.38(1H, m, 5″-H), 3.50 (2H, m, 6″-H) 2″-O-β-D-glucopyranosyl moiety; δ4.37(1H, d, J=7.8 Hz, 1′″-H), 3.03 (1H, dd, J=7.8, 9.0 Hz, 2′″-H), 3.17 (1H,t, J=9.0 Hz, 3′″-H), 3.07 (1H, t, J=9.0 Hz, 4′″-H), 3.15 (1H, m, 5′″-H),3.47 (1H, m,6′″-H), 3.73 (1H, m, 6′″-H) 22-O-α-L-arabinopyranosylmoiety; δ4.22 (1H, d, J=7.8 Hz, 1″″-H), 3.55 (1H, m, 2″″-H), 3.51 (1H,m, 3″″-H), 3.75 (1H, m, 4″″-H), 3.40 (1H, m, 5″″-H), 3.63 (1H, m, 5″″-H)3″″-O-β-D-xylopyranosyl moiety; δ4.47 (1H, m, 1″″′-H), 3.23 (1H,m,2″″′-H), 4.72 (1H, t, J=9.3 Hz, 3″″′-H), 3.48 (1H, m, 4″″′-H), 3.18(1H, m, 5″″′-H), 3.74 (1H, m, 5″″′-H), 2.03 (3H, s, 3″″′-CH₃)

Here, in ¹H-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of the residual proton of deuterateddimethyl sulfoxide was expressed as 2.51 ppm. FIG. 5 shows ¹H-NMRspectrum of the fraction 9 derived from the water extract of soybeanembryo. In FIG. 5, the axis of abscissas is chemical shift, and the axisof ordinates is intensity of signal.

¹³C-NMR: sapogenol moiety; δ 90.4 (3-C), 122.5 (12-C), 144.4 (13-C),75.0 (21-C), 92.3 (22-C), 63.0 (24-C) 3-O-β-D-glucronopyranosyl moiety;δ104.1 (1′-C), 78.9 (2′-C), 76.7 (3′-C), 72.0 (4′-C), 76.1 (5′-C), 171.0(6′-C) 2′-O-β-D-galactopyranosyl moiety; δ101.6 (1″-C), 82.7 (2″-C),73.4(3″-C), 69.0 (4″-C), 75.7 (5″-C), 61.0 (6″-C) 2″-O-β-D-glucopyranosylmoiety; δ105.3 (1′″-C), 75.5 (2′″-C), 76.8 (3′″-C), 70.6 (4′″-C), 78.1(5′″-C), 61.8 (6′″-C) 22-O-α-L-arabinopyranosyl moiety; δ107.4 (1″″-C),71.9 (2″″-C), 83.7 (3″″-C), 68.5 (4″″-C), 67.0 (5″″-C)3″″-O-β-D-xylopyranosyl moiety; δ105.6 (1″″′-C), 72.2 (2″″′-C), 77.8(3″″′-C), 68.1 (4″″′-C), 66.3 (5″″′-C), 170.8 (3″″′-C═O), 21.9(3″″′-CH₃)

Here, in ¹³C-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 40.2 ppm. FIG. 6 shows ¹³C-NMR spectrum of the fraction 9derived from the water extract of soybean embryo. In FIG. 6, the axis ofabscissas is chemical shift, and the axis of ordinates is intensity ofsignal.

The fraction 9 derived from the water extract of soybean embryo wassubjected to hydrolysis in the same manner as in item (1) of Example 2,to produce a saccharide component and a non-saccharide component. Thenon-saccharide component was subjected to thin layer chromatography(developing solvent A). As a result, a spot having the same Rf values asin Soyasapogenol A was detected. The saccharide component was subjectedto thin layer chromatography (developing solvent B). As a result,arabinose, glucose, xylose and galactose were detected.

From the above results, it was identified that the fraction 9 derivedfrom the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[3-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1280). The compound is a compound b in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound b wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound b has an enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.739 ng/mL.

Example 4 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[4-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The mass spectrum of the fraction 10 derived from the water extractof soybean embryo (fraction including a peak detected at a retentiontime of 35.6 minutes) fractionated in item (6) of Example 1 was measuredin the same manner as in item (1) of Example 2. As the matrix,triethanolamine was used. According to mass spectrometry, a peak of m/z1279 (M−H)⁻ was detected. FIG. 7 shows the mass spectrum of the fraction10 derived from the water extract of soybean embryo. In FIG. 7, the axisof abscissas is m/z value, and the axis of ordinates is relativeintensity.

The NMR spectrum of the fraction 10 derived from the water extract ofsoybean embryo was measured in the same manner as in item (1) of Example2. The signals of NMR are shown below.

¹H-NMR: sapogenol moiety; δ3.26 (1H, m, 3-H), 5.15 (1H, br-s, 12-H),3.39 (1H, m, 21-H), 3.23 (1H, br-s, 22-H), 3.15 (1H, m, 24-H), 3.86 (1H,m, 24-H) 3-O-β-D-glucronopyranosyl moiety; δ4.46 (1H, d, J=7.2 Hz,1′-H), 3.43 (1H, m, 2′-H), 3.56 (1H, m, 3′-H), 3.33 (1H, m, 4′-H), 3.65(1H, m, 5′-H) 2′-O-β-D-galactopyranosyl moiety; δ4.81 (1H, d, J=7.2 Hz,1″-H), 3.51 (1H, m, 2″-H), 3.51 (1H, m, 3″-H), 3.65 (1H, m, 4″-H), 3.38(1H, m, 5″-H), 3.50 (2H, m, 6″-H) 2″-O-β-D-glucopyranosyl moiety; δ4.38(1H, d, J=7.8 Hz, 1′″-H), 3.02 (1H, m, 2′″-H), 3.16 (1H, m, 3′″-H), 3.08(1H, m, 4′″-H), 3.15 (1H, m, 5′″-H), 3.47 (1H, m, 6′″-H), 3.73 (1H, m,6′″-H) 22-O-α-L-arabinopyranosyl moiety; δ4.22 (1H, d, J=7.2 Hz, 1″″-H),3.57 (1H, m, 2″″-H), 3.46 (1H, m, 3″″-H), 3.74 (1H, m, 4″″-H), 3.39 (1H,m, 5″″-H), 3.62 (1H, m, 5″″-H) 3″″-O-β-D-xylopyranosyl moiety; δ4.43(1H, d, J=7.2 Hz, 1″″′-H), 3.39 (1H, m, 2″″′-H), 3.18 (1H, m, 3″″′-H),4.53 (1H, m, 4″″′-H), 3.17 (1H, m, 5″″′-H), 3.78 (1H, m, 5″″′-H), 2.01(3H, s, 4″″′-CH₃)

Here, in ¹H-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 2.51 ppm. FIG. 8 shows ¹H-NMR spectrum of the fraction 10derived from the water extract of soybean embryo. In FIG. 8, the axis ofabscissas is chemical shift, and the axis of ordinates is intensity ofsignal.

¹³C-NMR: sapogenol moiety; δ 90.3 (3-C), 122.5 (12-C), 144.4 (13-C),75.0 (21-C), 92.4 (22-C), 63.0 (24-C) 3-O-β-D-glucronopyranosyl moiety;δ104.1 (1′-C), 78.8 (2′-C), 76.5 (3′-C), 71.9 (4′-C), 76.2 (5′-C), 171.0(6′-C) 2′-O-β-D-galactopyranosyl moiety; δ101.5 (1″-C), 82.7 (2″-C),73.3 (3″-C), 69.0 (4″-C), 75.7 (5″-C), 60.9 (6″-C)2″-O-β-D-glucopyranosyl moiety; δ105.3 (1′″-C), 75.5 (2′″-C), 76.8(3′″-C), 70.6 (4′″-C), 78.1 (5′″-C), 61.8 (6′″-C)22-O-α-L-arabinopyranosyl moiety; δ107.5 (1″″-C), 71.8 (2″″-C), 83.9(3″″-C), 68.4 (4″″-C), 67.0 (5″″-C) 3″″-O-β-D-xylopyranosyl moiety;δ105.9 (1″″′-C), 73.3 (2″″′-C), 74.5 (3″″′-C), 72.3 (4″″′-C), 62.8(5″″′-C), 171.0 (4″″′-C═O), 21.7 (4″″′-CH₃)

Here, in ¹³C-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 40.2 ppm. FIG. 9 shows ¹³C-NMR spectrum of the fraction 10derived from the water extract of soybean embryo. In FIG. 9, the axis ofabscissas is chemical shift, and the axis of ordinates is intensity ofsignal.

The fraction 10 derived from the water extract of soybean embryo wassubjected to hydrolysis in the same manner as in item (1) of Example 2,to produce a saccharide component and a non-saccharide component. Thenon-saccharide component was subjected to thin layer chromatography(developing solvent A). As a result, a spot having the same Rf values asin Soyasapogenol A was detected. The saccharide component was subjectedto thin layer chromatography (developing solvent B). As a result,arabinose, glucose, xylose and galactose were detected.

From the above results, it was identified that the fraction 10 derivedfrom the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[4-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1280). The compound is a compound c in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound c wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound c has an enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.739 ng/mL.

Example 5 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,3-di-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]sovasapogenolA

(1) The mass spectrum of the fraction 12 derived from the water extractof soybean embryo (fraction including a peak detected at a retentiontime of 38.2 minutes) fractionated in item (6) of Example 1 was measuredin the same manner as in item (1) of Example 2. As the matrix,m-nitrobenzyl alcohol was used. According to mass spectrometry, a peakof m/z 1321(M−H)⁻ was detected. FIG. 10 shows the mass spectrum of thefraction 12 derived from the water extract of soybean embryo. In FIG.10, the axis of abscissas is m/z value, and the axis of ordinates isrelative intensity.

The NMR spectrum of the fraction 12 derived from the water extract ofsoybean embryo was measured in the same manner as in item (1) of Example2. The signals of NMR are shown below.

¹H-NMR: sapogenol moiety; δ3.26(1H, m, 3-H), 5.14(1H, br-s, 12-H), 3.43(1H, m, 21-H), 3.23 (1H, br-s, 22-H), 3.15 (1H, m, 24-H), 3.86 (1H, m,24-H) 3-O-β-D-glucronopyranosyl moiety; δ4.46(1H, d, J=7.8 Hz, 1′-H),3.44 (1H, m, 2′-H), 3.56 (1H, m, 3′-H), 3.33 (1H, m, 4′-H), 3.65 (1H, m,5′-H) 2′-O-β-D-galactopyranosyl moiety; δ4.80 (1H, d, J=6.6 Hz, 1″-H),3.50 (1H, m, 2″-H), 3.51 (1H, m, 3″-H), 3.65 (1H, m, 4″-H), 3.38 (1H, m,5″-H), 3.50 (2H, m, 6″-H) 2″-O-β-D-glucopyranosyl moiety; δ4.37 (1H, d,J=7.8 Hz, 1′″-H), 3.03 (1H, m, 2′″-H), 3.17 (1H, m, 3′″-H), 3.08 (1H, m,4′″-H), 3.14 (1H, m, 5′″-H), 3.47 (1H, m, 6′″-H), 3.73 (1H, m, 6′″-H)22-O-α-L-arabinopyranosyl moiety; δ4.15 (1H, d, J=6.6 Hz, 1″″-H), 3.42(1H, m, 2″″-H), 3.45 (1H, m, 3″″-H), 3.71 (1H, m, 4″″-H), 3.35 (1H, m,5″″-H), 3.62 (1H, m, 5″″-H) 3″″-O-β-D-xylopyranosyl moiety; δ4.74 (1H,d, J=7.2 Hz, 1″″′-H), 4.65 (1H, m, 2″″′-H), 4.83 (1H, t, J=7.2 Hz,3″″′-H), 3.60 (1H, m, 4″″′-H), 3.82 (1H, m, 5″″′-H), 3.24 (1H, m,5″″′-H), 1.95 (3H, s, 2″″′-CH₃), 1.98 (3H, s, 3″″′-CH₃)

Here, in ¹H-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of the residual proton of deuterateddimethyl sulfoxide was expressed as 2.51 ppm. FIG. 11 shows ¹H-NMRspectrum of the fraction 12 derived from the water extract of soybeanembryo. In FIG. 11, the axis of abscissas is chemical shift, and theaxis of ordinates is intensity of signal.

¹³C-NMR: sapogenol moiety; δ 90.3 (3-C), 122.5 (12-C), 144.4 (13-C),74.8 (21-C), 91.8 (22-C), 63.0 (24-C) 3-O-β-D-glucronopyranosyl moiety;δ104.1 (1′-C), 79.0 (2′-C), 76.6 (3′-C), 71.9 (4′-C), 76.1 (5′-C), 171.0(6′-C) 2′-O-β-D-galactopyranosyl moiety; δ101.5 (1″-C), 82.7 (2″-C),73.3 (3″-C), 69.0 (4″-C), 75.8 (5″-C), 60.9 (6″-C)2″-O-β-D-glucopyranosyl moiety; δ105.3 (1′″-C), 75.5 (2′″-C), 76.7(3′″-C), 70.6 (4′″-C), 78.1 (5′″-C), 61.8 (6′″-C)22-O-α-L-arabinopyranosyl moiety; δ107.9 (1″″′-C), 72.0(2″″′-C), 83.5(3″″′-C), 68.4 (4″″′-C), 67.0 (5″″′-C) 3″″-O-β-D-xylopyranosyl moiety;δ102.8 (1″″′-C), 72.4 (2″″′-C), 76.0 (3″″′-C), 68.0 (4″″′-C), 66.0(5″″′-C), 170.4 (2″″″-C═O), 21.5 (2″″′-CH₃), 170.7 (3″″′-C═O), 21.5(3″″′-CH₃)

Here, in ¹³C-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 40.2 ppm. FIG. 12 shows ¹³C-NMR spectrum of the fraction 12derived from the water extract of soybean embryo. In FIG. 12, the axisof abscissas is chemical shift, and the axis of ordinates is intensityof signal.

The fraction 12 derived from the water extract of soybean embryo wassubjected to hydrolysis in the same manner as in item (1) of Example 2,to produce a saccharide component and a non-saccharide component. Thenon-saccharide component was subjected to thin layer chromatography(developing solvent A). As a result, a spot having the same Rf values asin Soyasapogenol A was detected. The saccharide component was subjectedto thin layer chromatography (developing solvent B). As a result,arabinose, glucose, xylose and galactose were detected.

From the above results, it was identified that the fraction 12 derivedfrom the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,3-di-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1322). The compound is a compound d in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound d wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound d has an enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.739 ng/mL.

Example 6 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,4-di-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The mass spectrum of the fraction 13 derived from the water extractof soybean embryo (fraction including a peak detected at a retentiontime of 38.9 minutes) fractionated in item (6) of Example 1 was measuredin the same manner as in item (1) of Example 2. As the matrix,m-nitrobenzyl alcohol was used. According to mass spectrometry, a peakof m/z 1321(M−H)⁻ was detected. FIG. 13 shows the mass spectrum of thefraction 13 derived from the water extract of soybean embryo. In FIG.13, the axis of abscissas is m/z value, and the axis of ordinates isrelative intensity.

The NMR spectrum of the fraction 13 derived from the water extract ofsoybean embryo was measured in the same manner as in item (1) of Example2. The signals of NMR are shown below.

¹H-NMR: sapogenol moiety; δ3.27 (1H, m, 3-H), 5.14 (1H, br-s, 12-H),3.45 (1H, m, 21-H), 3.22 (1H, br-s, 22-H), 3.12 (1H, m, 24-H), 3.85 (1H,m, 24-H) 3-O-β-D-glucronopyranosyl moiety; δ4.46 (1H, d, J=7.2 Hz,1′-H), 3.43 (1H, m, 2′-H), 3.57 (1H, m, 3′-H), 3.33 (1H, m, 4′-H), 3.64(1H, m, 5′-H) 2′-O-β-D-galactopyranosyl moiety; δ4.81 (1H, d, J=6.6 Hz,1″-H), 3.51 (1H, m, 2″-H), 3.51 (1H, m, 3″-H), 3.64 (1H, m, 4″-H), 3.38(1H, m, 5″-H), 3.50 (2H, m, 6″-H) 2″-O-β-D-glucopyranosyl moiety; δ4.38(1H, d, J=7.8 Hz, 1′″-H), 3.02 (1H, m, 2′″-H), 3.17 (1H, m, 3′″-H), 3.08(1H, m, 4′″-H), 3.14 (1H, m, 5′″-H), 3.47 (1H, m, 6′″-H), 3.72 (1H, m,6′″-H) 22-O-α-L-arabinopyranosyl moiety; δ4.15 (1H, d, 1″″-H), 3.44 (1H,m, 2″″-H), 3.43 (1H, m, 3″″-H), 3.69 (1H, m, 4″″-H), 3.35 (1H, m,5″″-H), 3.60 (1H, m, 5″″-H) 3″″-O-β-D-xylopyranosyl moiety; δ4.70 (1H,d, J=7.2 Hz, 1″″′-H), 4.62 (1H, m, 2″″′-H), 3.60 (1H, m, 3″″′-H), 4.59(1H, m, 4″″′-H), 3.27 (1H, m, 5″″′-H), 3.90 (1H, m, 5″″′-H), 2.02 (3H,s, 2″″′-CH₃), 2.02 (3H, s, 4″″′-CH₃)

Here, in ¹H-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of the residual proton of deuterateddimethyl sulfoxide was expressed as 2.51 ppm. FIG. 14 shows ¹H-NMRspectrum of the fraction 13 derived from the water extract of soybeanembryo. In FIG. 14, the axis of abscissas is chemical shift, and theaxis of ordinates is intensity of signal.

¹³C-NMR: sapogenol moiety; δ90.3 (3-C), 122.5 (12-C), 144.4 (13-C), 74.8(21-C), 91.8 (22-C), 63.0 (24-C) 3-O-β-D-glucronopyranosyl moiety;δ104.1 (1′-C), 78.7 (2′-C), 76.6 (3′-C), 71.9 (4′-C), 76.1 (5′-C), 171.0(6′-C) 2′-O-β-D-galactopyranosyl moiety; δ101.5 (1″-C), 82.7 (2″-C),73.3 (3″-C), 69.0 (4″-C), 75.5 (5″-C), 60.9 (6″-C)2″-O-β-D-glucopyranosyl moiety; δ105.3 (1′″-C), 75.5 (2′″-C), 76.7(3′″-C), 70.6 (4′″-C), 78.1 (5′″-C), 61.8 (6′″-C)22-O-α-L-arabinopyranosyl moiety; δ108.0 (1″″-C), 71.9 (2″″-C), 83.0(3″″-C), 68.2 (4″″-C), 67.0 (5″″-C) 3″″-O-β-D-xylopyranosyl moiety;δ102.6 (1″″′-C), 73.7 (2″″′-C), 70.6 (3″″′-C), 71.6 (4″″′-C), 62.0(5″″′-C), 170.4, 170.9 (2″″′ and 4″″′-C═O), 21.6, 21.8 (2″″′ and4″″′-CH₃)

Here, in ¹³C-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 40.2 ppm. FIG. 15 shows ¹³C-NMR spectrum of the fraction 13derived from the water extract of soybean embryo. In FIG. 15, the axisof abscissas is chemical shift, and the axis of ordinates is intensityof signal.

The fraction 13 derived from the water extract of soybean embryo wassubjected to hydrolysis in the same manner as in item (1) of Example 2,to produce a saccharide component and a non-saccharide component. Thenon-saccharide component was subjected to thin layer chromatography(developing solvent A). As a result, a spot having the same Rf values asin Soyasapogenol A was detected. The saccharide component was subjectedto thin layer chromatography (developing solvent B). As a result,arabinose, glucose, xylose and galactose were detected.

From the above results, it was identified that the fraction 13 derivedfrom the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,4-di-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1322). The compound is a compound e in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound e wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound e has an enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.578 ng/mL.

Example 7 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[3,4-di-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The mass spectrum of the fraction 15 derived from the water extractof soybean embryo (fraction including a peak detected at a retentiontime of 40.3 minutes) fractionated in item (6) of Example 1 was measuredin the same manner as in item (1) of Example 2. As the matrix,m-nitrobenzyl alcohol was used. According to mass spectrometry, a peakof m/z 1321(M−H)⁻ was detected. FIG. 16 shows the mass spectrum of thefraction 15 derived from the water extract of soybean embryo. In FIG.16, the axis of abscissas is m/z value, and the axis of ordinates isrelative intensity.

The NMR spectrum of the fraction 15 derived from the water extract ofsoybean embryo was measured in the same manner as in item (1) of Example2. The signals of NMR are shown below.

¹H-NMR: sapogenol moiety; δ3.27 (1H, m, 3-H), 5.14 (1H, br-s, 12-H),3.39 (1H, m, 21-H), 3.24 (1H, br-s, 22-H), 3.14 (1H, m, 24-H), 3.85 (1H,m, 24-H) 3-O-β-D-glucronopyranosyl moiety; δ4.45 (1H, d, J=7.8 Hz,1′-H), 3.43 (1H, m, 2′-H), 3.56 (1H, t, J=8.9 Hz, 3′-H), 3.34 (1H, t,J=8.9 Hz, 4′-H), 3.66 (1H, m, 5′-H) 2′-O-β-D-galactopyranosyl moiety;δ4.80 (1H, d, J=6.6 Hz, 1″-H), 3.51 (1H, m, 2″-H), 3.51 (1H, m, 3″-H),3.65 (1H, m, 4″-H), 3.38 (1H, m, 5″-H), 3.50 (2H, m, 6″-H)2″-O-β-D-glucopyranosyl moiety; δ4.37 (1H, d, J=7.8 Hz, 1′″-H), 3.02(1H, dd, J=7.8, 9.0 Hz, 2′″-H), 3.17 (1H, t, J=9.0 Hz, 3′″-H), 3.08 (1H,t, J=9.0 Hz, 4′″-H), 3.15 (1H, m, 5′″-H), 3.47 (1H, m, 6′″-H), 3.72 (1H,m, 6′″-H) 22-O-α-L-arabinopyranosyl moiety; δ4.22 (1H, d, J=4.8 Hz,1″″-H), 3.46 (1H, m, 2″″-H), 3.42 (1H, m, 3″″-H), 3.74 (1H, m, 4″″-H),3.39 (1H, m, 5″″-H), 3.63 (1H, m, 5″″-H) 3″″-O-β-D-xylopyranosyl moiety;δ4.58 (1H, d, J=7.2 Hz, 1″″′-H), 3.38 (1H, m, 2″″′-H), 4.97 (1H, t,J=9.3 Hz, 3″″′-H), 4.72 (1H, m, 4″″′-H), 3.88 (1H, m, 5″″′-H), 3.36 (1H,m, 5″″′-H), 2.01, 1.96 (3H, s, 3″″′ and 4″″′-CH₃)

Here, in ¹H-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of the residual proton of deuterateddimethyl sulfoxide was expressed as 2.51 ppm. FIG. 17 shows ¹H-NMRspectrum of the reaction 15 derived from the water extract of soybeanembryo. In FIG. 17, the axis of abscissas is chemical shift, and theaxis of ordinates is intensity of signal.

¹³C-NMR: δ sapogenol moiety; δ90.3 (3-C), 122.5 (12-C), 144.4 (13-C),75.0 (21-C), 92.3 (22-C), 62.6 (24-C) 3-O-β-D-glucronopyranosyl moiety;δ104.1 (1′-C), 78.8 (2′-C), 76.5 (3′-C), 72.0 (4′-C), 76.1 (5′-C), 171.0(6′-C) 2′-O-β-D-galactopyranosyl moiety; δ101.5 (1″-C), 82.7(2″-C), 73.4(3″-C), 69.0 (4″-C), 75.5(5″-C), 60.9(6″-C) 2″-O-β-D-glucopyranosylmoiety; δ105.3(1′″-C), 75.5(2′″-C), 76.5 (3′″-C), 70.6 (4′″-C), 78.1(5′″-C), 61.8(6′″-C) 22-O-α-L-arabinopyranosyl moiety; δ107.4 (1″″-C),72.0 (2″″-C), 83.7 (3″″-C), 68.3 (4″″-C), 67.0(5″″-C)3″″-O-β-D-xylopyranosyl moiety; δ105.3 (1″″′-C), 72.0 (2″″′-C), 74.2(3″″′-C), 69.9 (4″″′-C), 62.6 (5″″′-C), 171.0, 170.7 (3″″′ and4″″′-C═O), 21.3, 21.5 (3″″′ and 4″″′-CH₃)

Here, in ¹³C-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 40.2 ppm. FIG. 18 shows ¹³C-NMR spectrum of the fraction 15derived from the water extract of soybean embryo. In FIG. 18, the axisof abscissas is chemical shift, and the axis of ordinates is intensityof signal.

The fraction 15 derived from the water extract of soybean embryo wassubjected to hydrolysis in the same manner as in item (1) of Example 2,to produce a saccharide component and a non-saccharide component. Thenon-saccharide component was subjected to thin layer chromatography(developing solvent A). As a result, a spot having the same Rf values asin Soyasapogenol A was detected. The saccharide component was subjectedto thin layer chromatography (developing solvent B). As a result,arabinose, glucose, xylose and galactose were detected.

From the above results, it was identified that the fraction 15 derivedfrom the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[3,4-di-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1322). The compound is a compound f in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound f wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound f has an enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.578 ng/mL.

Example 8 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[3,4,6-tri-O-acetyl-α-D-glucopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The mass spectrum of the fraction 16 derived from the water extractof soybean embryo (fraction including a peak detected at a retentiontime of 41.0 minutes) fractionated in item (6) of Example 1 was measuredin the same manner as in item (1) of Example 2. As the matrix,m-nitrobenzyl alcohol was used. According to mass spectrometry, a peakof m/z 1393 (M−H)⁻ was detected. FIG. 19 shows the mass spectrum of thefraction 16 derived from the water extract of soybean embryo. In FIG.19, the axis of abscissas is m/z value, and the axis of ordinates isrelative intensity.

The NMR spectrum of the fraction 16 derived from the water extract ofsoybean embryo was measured in the same manner as in item (1) of Example2. The signals of NMR are shown below.

¹H-NMR: sapogenol moiety; δ3.28 (1H, m, 3-H), 5.15 (1H, br-s, 12-H),3.40 (1H, m, 21-H), 3.25 (1H, br-s, 22-H), 3.15 (1H, m, 24-H), 3.87 (1H,m, 24-H) 3-O-β-D-glucronopyranosyl moiety; δ4.45 (1H, d, 1′-H), 3.44(1H, m,2′-H), 3.57 (1H, m, 3′-H), 3.32 (1H, m, 4′-H), 3.62 (1H, m, 5′-H)2′-O-β-D-galactopyranosyl moiety; δ4.82 (1H, d, 1″-H), 3.51 (1H, m,2″-H), 3.51 (1H, m, 3″-H), 3.65 (1H, m, 4″-H), 3.38 (1H, m, 5″-H), 3.50(2H, m, 6″-H) 2″-O-β-D-glucopyranosyl moiety; δ4.38 (1H, d, J=7.8 Hz,1′″-H), 3.02 (1H, m, 2′″-H), 3.17 (1H, m, 3′″-H), 3.09 (1H, m, 4′″-H),3.14 (1H, m, 5′″-H), 3.47 (1H, m, 6′″-H), 3.72 (1H, m, 6′″-H)22-O-α-L-arabinopyranosyl moiety; δ4.25 (1H, d, 1″″-H), 3.59 (1H, m,2″″-H), 3.52 (1H, m, 3″″-H), 3.78 (1H, m, 4″″-H), 3.42 (1H, m, 5″″-H),3.66 (1H, m, 5″″-H) 3″″-O-β-D-glucopyranosyl moiety; δ4.67 (1H, d, J=7.8Hz, 1″″′-H), 3.41 (1H, dd, J=7.8, 9.3 Hz, 2″″′-H), 5.03 (1H, t, J=9.3Hz, 3″″′-H), 4.77 (1H, t, J=9.3 Hz, 4″″′-H), 3.87 (1H, m, 5″″′-H) 3.97(1H, m, 6″″′-H), 4.17 (1H, m, 6″″′-H), 1.96, 1.99 (3H and 6H, s,3″″′,4″″′ and 6″″′-CH₃)

Here, in ¹H-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of the residual proton of deuterateddimethyl sulfoxide was expressed as 2.51 ppm. FIG. 20 shows ¹H-NMRspectrum of the fraction 16 derived from the water extract of soybeanembryo. In FIG. 20, the axis of abscissas is chemical shift, and theaxis of ordinates is intensity of signal.

¹³C-NMR: sapogenol moiety; δ 90.3 (3-C), 122.5 (12-C), 144.4 (13-C),75.0 (21-C), 92.4 (22-C), 63.0 (24-C) 3-O-β-D-glucronopyranosyl moiety;δ104.1 (1′-C), 78.8 (2′-C), 76.6 (3′-C), 72.0 (4′-C), 76.0 (5′-C), 171.0(6′-C) 2′-O-β-D-galactopyranosyl moiety; δ101.5 (1″-C), 82.8 (2″-C),73.2 (3″-C), 69.0 (4″-C), 75.7 (5″-C), 60.9 (6″-C)2″-O-β-D-glucopyranosyl moiety; δ105.3 (1′″-C), 75.5 (2′″-C), 76.7(3′″-C), 70.6 (4′″-C), 78.1 (5′″-C), 61.8 (6′″-C)22-O-α-L-arabinopyranosyl moiety; δ107.4 (1″″-C), 71.6 (2″″-C), 84.4(3″″-C), 68.3 (4″″-C), 67.0 (5″″-C) 3″″-O-β-D-glucopyranosyl moiety;δ104.7 (1″″′-C), 72.2 (2″″′-C), 75.0 (3″″′-C), 69.4 (4″″′-C),71.3(5″″′-C), 62.8 (6″″′-C), 170.3, 170.5, 171.1 (3″″′, 4″″′ and6″″′-C═O), 21.3, 21.4, 21.5 (3″″′, 4″″′ and 6″″′-CH₃)

Here, in ¹³C-NMR, the sample was dissolved in deuterated dimethylsulfoxide, and the chemical shift of deuterated dimethyl sulfoxide wasexpressed as 40.2 ppm. FIG. 21 shows ¹³C-NMR spectrum of the fraction 16derived from the water extract of soybean embryo. In FIG. 21, the axisof abscissas is chemical shift, and the axis of ordinates is intensityof signal.

The fraction 16 derived from the water extract of soybean embryo wassubjected to hydrolysis in the same manner as in item (1) of Example 2,to produce a saccharide component and a non-saccharide component. Thenon-saccharide component was subjected to thin layer chromatography(developing solvent A). As a result, a spot having the same Rf values asin Soyasapogenol A was detected. The saccharide component was subjectedto thin layer chromatography (developing solvent B). As a result,arabinose, glucose, xylose and galactose were detected.

From the above results, it was identified that the fraction 16 derivedfrom the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[3,4,6-tri-O-acetyl-β-D-glucopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1394). The compound is a compound g in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound g wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound g has an enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.739 ng/mL.

Example 9 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The mass spectrum and NMR spectrum of the fraction 18 derived fromwater extract of soybean embryo (fraction including a peak detected at aretention time of 42.7 minutes) fractionated in item (6) of Example 1,and thin layer chromatography of the sacchatride moiety and thenon-saccharide moiety were measured in the same manner as in item (1) ofExample 2. As a result, it was identified that the fraction 18 derivedfrom the water extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1436). The compound is a compound h in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound h wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound h has enhancing activity for NGFproduction. The results are shown in Table 2. Incidentally, the amountof NGF production of the control was 0.898 ng/mL.

Example 10 Fractionation of Fraction Derived from 70% Ethanol Extract ofSoybean Embryo

(1) To the precipitate 2 (5.6 kg) obtained in item (1) of Example 1 wasadded 8.4 liters of ethanol, and the mixture was stirred at roomtemperature for 1 hour. Thereafter, centrifugation was carried out at5000 g for 10 minutes, to give a precipitate 6 and supernatant 6. Tenliters of 70% ethanol was added to the precipitate 6, and the mixturewas stirred for 1 hour. Thereafter, centrifugation was carried out at5000 g for 10 minutes, to give a precipitate 7 and supernatant 7. Thesupernatant 6 and the supernatant 7 were mixed and concentrated underreduced pressure, to give a concentrate of 70% ethanol extract ofsoybean embryo.

(2) The concentrate of 70% ethanol extract of soybean embryo obtained initem (1) of Example 10 was dissolved in 500 mL of 70% ethanol.Thereafter, centrifugation was carried out at 5000 g for 10 minutes, togive a precipitate 8 and supernatant 8. The supernatant 8 wasconcentrated under reduced pressure, and thereafter the concentrate wassuspended in 1000 mL of distilled water. The residue after filtrationwas obtained as a water-insoluble fraction derived from 70% ethanolextract of soybean embryo.

(3) In a solvent composed of 2 mL of methanol and 10 mL of chloroformwas dissolved 0.4 volumes of the water-insoluble fraction derived from70% ethanol extract of soybean embryo obtained in item (2) of Example10, and the mixture was fractionated using a silica column. Theconditions therefor are given below. BW-300SP (the amount of resin: 300mL) was used as silica gel. The elution was carried out usingchloroform:methanol:acetic acid=100:20:1 (400 mL),chloroform:methanol=2:1 (400 mL) and ethanol: water=10:1(200 mL), inthat order, as developing solvents. Fractions were collected for every100 mL portion, to give fractions 1 to 10. Each of the eluted fractionswas concentrated under reduced pressure, to give silica column fractions1 to 10 derived from the 70% ethanol extract.

(4) The silica column fractions 4 and 5 derived form the 70% ethanolextract derived in item (3) of Example 10 were combined andconcentrated, and the concentrate was then dissolved in 12 mL ofethanol. Thereafter, the mixture was fractionated using reverse phasechromatography. The conditions therefor are given below. TSK gelODS-80Ts (21.5 mm×30 cm) was used as the column. The elution ratio ofSolvent A (mixture prepared by mixing distilled water and acetonitrilein a volume ratio of 3:1) to Solvent B (mixture prepared by mixingdistilled water and acetonitrile in a volume ratio of 1:3) was such thatthe ratio of Solvent B increased linearly at 40 to 70% from 0 to 25minutes, the ratio of Solvent B increased linearly at 70 to 100% in thenext 25 to 26 minutes, and the ratio of Solvent B was retained at 100%during the subsequent 5 minutes. The elution rate was 5 mL/minute, andthe detection was carried out at 215 nm. Fractions 1 to 10 derived fromthe 70% ethanol extract of soybean embryo were fractionated usingultraviolet light absorption of the eluate as an index.

Example 11 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolB

(1) The mass spectrum and NMR spectrum of the fraction 6 derived fromthe 70% ethanol extract of soybean embryo (fraction including a peakdetected at a retention time of 24.8 minutes) fractionated in item (4)of Example 10 were measured in the same manner as in item (1) of Example2. Also, the fraction 6 derived from the 70% ethanol extract of soybeanembryo was heated in a mixed solution of 2 M trifluoroacetic acid: 2NHCl (1:1) in a gas phase at 100° C. for 4 hours, to produce a saccharidecomponent and a non-saccharide component. The non-saccharide moiety wassubjected to thin layer chromatography (developing solvent A). As aresult, a spot having the same Rf value as Soyasapogenol B was detected.The saccharide component was subjected to thin layer chromatography(developing solvent B). As a result, glucose and galactose weredetected. Consequently, it was identified that the fraction 6 derivedfrom the 70% ethanol extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolB (molecular weight: 958). The compound is a compound i in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound i wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound i has enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.948 ng/mL.

Example 12 Enhancing Activity for NGF Production of3-O-[α-L-rhamnopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolB

(1) The mass spectrum and NMR spectrum of the fraction 7 derived fromthe 70% ethanol extract of soybean embryo (fraction including a peakdetected at a retention time of 27.4 minutes) fractionated in item (4)of Example 10, and thin layer chromatography of the saccharide moietyand the non-saccharide moiety were measured in the same manner as initem (1) of Example 11. As a result, it was identified that the fraction7 derived from the 70% ethanol extract of soybean embryo is3-O-[α-L-rhamnopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolB (molecular weight: 942). The compound is a compound j in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound j wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound j has enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.948 ng/mL.

Example 13 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolE

(1) The mass spectrum and NMR spectrum of the fraction 8 derived fromthe 70% ethanol extract of soybean embryo (fraction including a peakdetected at a retention time of 29.3 minutes) fractionated in item (4)of Example 10, and thin layer chromatography of the saccharide moietyand the non-saccharide moiety were measured in the same manner as initem (1) of Example 11. As a result, it was identified that the fraction8 derived from the 70% ethanol extract of soybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolE (molecular weight: 956). The compound is a compound k in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound k wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound k has enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.924 ng/mL.

Example 14 Enhancing Activity for NGF Production of3-O-[α-L-rhamnopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolE

(1) The mass spectrum and NMR spectrum of the fraction 9 derived fromthe 70% ethanol extract of soybean embryo (fraction including a peakdetected at a retention time of 31.7 minutes) fractionated in item (4)of Example 10, and thin layer chromatography of the saccharide moietyand the non-saccharide moiety were measured in the same manner as initem (1) of Example 11. As a result, it was identified that the fraction9 derived from the 70% ethanol extract of soybean embryo is3-O-[α-L-rhamnopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-soyasapogenolE (molecular weight: 940). The compound is a compound l in Table 1mentioned above.

(2) The enhancing activity for NGF production of the compound l wasmeasured in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound l has enhancing activity for NGFproduction. The results are shown in Table 2. The amount of NGFproduction of the control was 0.924 ng/mL.

Example 15 Enhancing Activity for NGF Production of3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,3,4-tri-O-acetyl-β-D-xylopvranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA

(1) The silica column fraction 3 derived from the 70% ethanol extract ofsoybean embryo obtained in item (3) of Example 10 was dissolved in 5 mLof ethanol, and the mixture was fractionated with reverse phasechromatography. The conditions therefor are given below. TSK gelODS-80Ts (21.5 mm×30 cm) was used as the column. The elution ratio ofSolvent A (mixture prepared by mixing distilled water and acetonitrilein a volume ratio of 3:1) to Solvent B (mixture prepared by mixingdistilled water and acetonitrile in a volume ratio of 1:3) was such thatthe ratio of Solvent B was increased linearly at 40 to 45% from 0 to 15minutes, the ratio of Solvent B was increased linearly at 45 to 100%during the next 15 to 20 minutes, and the ratio of Solvent B wasretained at 100% during the subsequent 5 minutes. The elution rate was 5mL/minute, and the detection was carried out at 215 nm. Fractions werefractionated using ultraviolet light absorption of the eluate as anindex.

(2) The mass spectrum and NMR spectrum of the fraction A derived fromthe 70% ethanol extract of soybean embryo (fraction including a peakdetected at a retention time of 19.5 minutes) obtained by fractionatingthe silica column fraction 3 derived from the 70% ethanol extract ofsoybean embryo in item (1) of Example 15, and thin layer chromatographyof the saccharide moiety and the non-saccharide moiety were measured inthe same manner as in item (1) of Example 11. As a result, it wasidentified that the fraction A derived from the 70% ethanol extract ofsoybean embryo is3-O-[β-D-glucopyranosyl(1→2)-β-D-galactopyranosyl-(1→2)-β-D-glucuronopyranosyl]-22-O-[2,3,4-tri-O-acetyl-β-D-xylopyranosyl(1→3)-α-L-arabinopyranosyl]soyasapogenolA (molecular weight: 1364). The compound is a compound m in Table 1mentioned above.

(3) The enhancing activity for NGF production of the compound m wasassayed in the same manner as in item (2) of Example 2. As a result, itwas clarified that the compound m has enhancing activity for NGFproduction. The results are shown in Table 2. In addition, the amount ofNGF production of the control was 0.898 ng/mL. TABLE 2 Compounds Amountof NGF Production (%) of the Present Concentration of Sample (mM)Invention 0 0.5 1.0 2 Compound a 100 228.3 231.6 212.6 Compound b 100220.9 251.2 272.3 Compound c 100 255.1 289.2 323.7 Compound d 100 255.8281.1 276.5 Compound e 100 214.0 233.7 230.1 Compound f 100 204.5 208.8218.5 Compound g 100 295.4 277.2 247.6 Compound h 100 273.3 301.7 307.2Compound i 100 N. T. 116.8 169.4 Compound j 100 138.2 176.4 212.5Compound k 100 N. T. 115.9 456.3 Compound l 100 N. T. 145.9 531.2Compound m 100 263.3 250.9 233.4The amounts of NGF production in the sample-added group are expressed as% when the amounts of NGF production in the sample-non-added group(control) are regarded as 100%.N. T. means that no test was carried out.

Example 16 Enhancing Activity for NGF Production of13(18)-oleanene-3,22,24-triol

(1) Under the conditions of 50% methanol, 2 N hydrochloric acid, washeated at 80° C. for 3 hours 0.04 volumes of the water-insolublefraction derived from the 70% ethanol extract of soybean embryo obtainedin item (2) of Example 10. Thereafter, the reaction solution wasextracted with an equivolume of diethyl ether for 3 times in total. Theether layer obtained was concentrated under reduced pressure, to givewhite powder. The resulting white powder was washed with hexane, to givean acid hydrolyzate of soybean embryo.

(2) The acid hydrolyzate of soybean embryo obtained in item (1) ofExample 16 was dissolved in chloroform, and the mixture was fractionatedwith silica chromatography. The conditions therefor are given below.BW-300SP (the amount of resin: 100 mL) was used as silica gel. Theelution was carried out using chloroform:methanol=100:1 as a developingsolvent, to give eluted fractions of a silica column fraction 1 derivedfrom the acid hydrolyzate of soybean embryo (550 mL), a silica columnfraction 2 derived from the acid hydrolyzate of soybean embryo (300 mL),and a silica column fraction 3 derived from the acid hydrolyzate ofsoybean embryo (250 mL) in that order.

(3) The silica column fraction 2 derived from the acid hydrolyzate ofsoybean embryo obtained in item (2) of Example 16 was dissolved in 4 mLof ethanol, and the mixture was fractionated using reverse phasechromatography. The conditions are given below. TSK gel ODS-80Ts (21.5mm×30 cm) was used as the column. A mixture prepared by mixing water andacetonitrile in a volume ratio of 1:9 was used as a solvent, the elutionrate was 5 mL/minute, and the detection was carried out at 215 nm.Fractions derived from the acid hydrolyzate of soybean embryo werefractionated using ultraviolet light absorption of the eluate as anindex.

(4) The mass spectrum of the fraction a derived from the acidhydrolyzate of soybean embryo fractionated in item (3) of Example 16(fraction including a peak detected at an elution time of 37.5 minutes)was measured in the same manner as in item (1) of Example 2.Triethanolamine was used as a matrix. According to mass spectrometry, apeak of m/z 457(M−H)⁻ was detected. The mass spectrum of the fraction aderived from the acid hydrolyzate of soybean embryo is shown in FIG. 22.In FIG. 22, the axis of abscissas is m/z value, and the axis ofordinates is relative intensity.

The NMR spectrum of the fraction a derived from the acid hydrolyzate ofsoybean embryo was measured in the same manner as in item (1) of Example2. The signals of NMR are shown below.

¹H-NMR: δ0.70, 0.92 (3H, s, 29 and 30-CH₃), 0.77 (1H, m, 5-H), 0.79 (3H,s, 26-CH₃), 0.81 (3H, s, 25-CH₃), 0.88 (3H, s, 28-CH₃), 0.96 (1H, m,1-H), 1.07 (1H, m, 15-H), 1.08 (3H, s, 23-CH₃), 1.12 (3H, s, 27-CH₃),1.16 (1H, dd, J=4.8, 13.8 Hz, 11-H), 1.25 (1H, m, 16-H), 1.27(1H, m,21-H), 1.32 (1H, m, 6-H), 1.34 (1H, m, 21-H), 1.34 (1H, m, 7-H), 1.37(1H, m, 7-H), 1.43 (1H, m, 9-H), 1.45 (1H, m, 11-H), 1.55 (1H, m, 6-H),1.56 (1H, m, 2-H), 1.61 (1H, m, 15-H), 1.62 (1H, m, 19-H), 1.64 (1H, m,2-H), 1.65 (1H, m, 1-H), 1.68 (1H, m, 16-H), 1.80 (1H, m, 12-H), 2.24(1H, d, J=13.8 Hz, 19-H), 2.61 (1H, br-d, J=12.6 Hz, 12-H), 3.12 (1H, m,22-H), 3.19 (1H, m, 3-H), 3.26 (1H, dd, J=7.8, 10.8 Hz, 24-H), 3.80 (1H,dd, J=2.4, 10.8 Hz, 24-H), 4.06 (1H, dd, J=2.4, 7.8 Hz, 24-OH)), 4.26(1H, d, J=4.8 Hz, 22-OH), 4.95 (1H, d, J=4.2 Hz, 3-OH)

The sample was dissolved in deuterated dimethyl sulfoxide in ¹H-NMR, andthe chemical shift of the residual proton of deuterated dimethylsulfoxide was shown as 2.51 ppm. FIG. 23 shows ¹H-NMR spectrum of thefraction a derived from the acid hydrolyzate of soybean embryo. In FIG.23, the axis of abscissas is the chemical shift, and the axis ofordinates is the intensity of signal.

¹³C-NMR: δ17.3 (25-C), 17.4 (28-C), 18.1 (26-C), 19.6 (6-C), 22.1(27-C), 22.4 (11-C), 23.8 (23-C), 25.8, 33.1 (29 and 30-C), 26.1 (12-C),26.6 (15-C), 28.2 (2-C), 32.7 (20-C), 34.0 (16-C), 35.8 (7-C), 37.5(10-C), 38.6 (19-C), 39.3 (1-C), 40.9 (17-C), 41.5 (8-C), 43.1 (4-C),44.7 (14-C), 44.8 (21-C), 51.0 (9-C), 56.3 (5-C), 63.8 (24-C), 76.6(22-C), 79.5 (3-C), 133.0 (18-C), 136.9 (13-C)

The sample was dissolved in deuterated dimethyl sulfoxide in ¹³C-NMR,and the chemical shift of deuterated dimethyl sulfoxide was shown as40.2 ppm. FIG. 24 shows ¹³C-NMR spectrum of the fraction a derived fromthe acid hydrolyzate of soybean embryo. In FIG. 24, the axis ofabscissas is the chemical shift, and the axis of ordinates is theintensity of signal.

As described above, as a result of the analyses of the MS spectrum andNMR spectrum for the fraction a derived from the acid hydrolyzate ofsoybean embryo, it was identified that the fraction a derived from theacid hydrolyzate of soybean embryo is 13(18)-oleanene-3,22,24-triol(molecular weight: 458). The compound is a compound n in Table 1mentioned above.

(5) The enhancing activity for NGF production of the compound n wasassayed in the same manner as in item (2) of Example 2. The amount addedto a medium was set so as to have a final concentration as shown inTable 3. As a result, it was clarified that the compound n has enhancingactivity for NGF production. The results are shown in Table 3. Theamount of NGF production of the control was 0.372 ng/mL. TABLE 3 Amountof NGF Production (%) Compound of the Concentration of Sample (mM)Present Invention 0 25 50 Compound n 100 197.9 522.6The amounts of NGF production in the sample-added group are expressed as% when the amounts of NGF production in the sample-non-added group(control) are regarded as 100%.

Example 17 Enhancing Activity for NGF Production of Glycyrrhizin

The enhancing activity for NGF production of glycyrrhizin (manufacturedby Wako Pure Chemical Industries) was assayed in the same manner as initem (2) of Example 2. The amount added to a medium was set so as tohave a final concentration as shown in Table 4. As a result, it wasclarified that glycyrrhizin has enhancing activity for NGF production.The results are shown in Table 4. The amount of NGF production of thecontrol was 0.484 ng/mL. TABLE 4 Compound of Amount of NGF Production(%) the present Concentration of Sample (mM) invention 0 0.25 0.5 1.0Glycyrrhizin 100 113.0 204.3 261.0The amounts of NGF production in the sample-added group are expressed as% when the amounts of NGF production in the sample-non-added group(control) are regarded as 100%.

INDUSTRIAL APPLICABILITY

According to the present invention, there is provided a medicament,foodstuff or feed, containing in a high content a saponin compound, asapogenin compound or a salt thereof which is derived from an edibleplant, and is safe. These are useful in treatment, prevention oramelioration of symptoms of cranial nerve-derived diseases by theirenhancing action for NGF production. Especially, the foodstuff canenable intake of a given amount of a saponin compound or the like on adaily basis, so that the foodstuff is very useful as a foodstuff formaintaining homeostasis of a living body or a foodstuff for enhancingNGF production.

1. A compound represented by the following general formula (I):

wherein of the bond lines indicated by dotted lines, when A is a doublebond and B is a single bond, R₁ is —O-GlcUA-Gal-Glc, R₂ is—O-Ara-2-AcXyl, —O-Ara-3-AcXyl, —O-Ara-4-AcXyl, —O-Ara-2,3-diAcXyl,—O-Ara-2,4-diAcXyl, —O-Ara-3,4-diAcXyl or —O-Ara-3,4,6-triAcGlc, and R₃is —OH; alternatively, when A is a single bond and B is a double bond,R₁ is —OH, R₂ is —OH and R₃ is —H, wherein GlcUA is glucuronic acidresidue, Gal is Galactose residue, Glc is glucose residue, Ara isarabinose residue, AcXyl is acetylated xylose residue, AcGlc isacetylated glucose residue, or a salt thereof:
 2. A therapeutic agent orprophylactic agent, characterized in that the therapeutic agent orprophylactic agent comprises as an effective ingredient the compound asdefined in claim 1 and/or a pharmacologically acceptable salt thereof,wherein the agent is for a disease showing sensitivity to the abovecompound.
 3. An enhancing agent for nerve growth factor production,characterized in that the enhancing agent comprises as an effectiveingredient the compound as defined in claim 1 and/or a salt thereof. 4.A food, beverage or feed, characterized in that the food, beverage orfeed comprises as an effective ingredient the compound as defined inclaim 1 and/or a salt thereof.
 5. The food, beverage or feed accordingto claim 4, wherein the food, beverage or feed is a food, beverage orfeed used for enhancing nerve growth factor production.
 6. A therapeuticagent or prophylactic agent for a disease requiring enhancement of nervegrowth factor production upon treatment or prevention, characterized inthat the therapeutic agent or prophylactic agent comprises as aneffective ingredient a compound selected from the group consisting ofsoyasaponin compounds, soyasapogenin compounds, glycyrrhizin andpharmacologically acceptable salts thereof.
 7. The therapeutic agent orprophylactic agent according to claim 6, wherein the soyasaponincompound and the soyasapogenin compound are a soyasaponin compound and asoyasapogenin compound which are represented by the following generalformula (II):

wherein the bond lines indicated by dotted lines are a single bond or adouble bond; R₄ is —OH or —O-saccharide residue; R₅ is —OH, ═O or—O-saccharide residue; and R₆ is —OH or —H.
 8. An enhancing agent fornerve growth factor production, characterized in that the enhancingagent comprises as an effective ingredient a compound selected from thegroup consisting of soyasaponin compounds, soyasapogenin compounds,glycyrrhizin and salts thereof.
 9. The enhancing agent according toclaim 8, wherein the soyasaponin compound and the soyasapogenin compoundare a soyasaponin compound and a soyasapogenin compound which arerepresented by the general formula (II) as defined in claim
 7. 10. Afood, beverage or feed for enhancing nerve growth factor production,characterized in that the food, beverage or feed comprises as aneffective ingredient a compound selected from the group consisting ofsoyasaponin compounds, soyasapogenin compounds, glycyrrhizin and saltsthereof.
 11. The food, beverage or feed according to claim 10, whereinthe soyasaponin compound and the soyasapogenin compound are asoyasaponin compound and a soyasapogenin compound which are representedby the general formula (II) as defined in claim 7.